Three adenine nucleotide binding sites in F1‐F0 mitochondrial ATPase as revealed by presteady‐state and steady‐state kinetics of ATP hydrolysis Evidence for two inhibitory ADP‐specific noncatalytic sites

FEBS Letters

Vinogradov, A.D.¹  Bulygin, V.V.¹ 

[1] Department of Biochemistry, School of Biology, Moscow State University, 119899 Moscow, USSR

关键词: F1-ATPase;    Steady-state ATP hydrolysis;    Nucleotide-binding site;    ClCCP;    carbonylcyanide m-chlorophenylhydrazone;   

DOI  :  10.1016/0014-5793(88)80085-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Preincubation of submitochondrial particles with ADP in the presence of Mg²⁺ results in the complete inhibition of ATPase which is slowly reactivated in the assay mixture containing ATP and the ATP regenerating system. Significantly, the rate of activation increases as the concentration of ADP in the preincubation mixture rises from 1 μM to 20 μM and reaches a constant value at higher ADP concentrations. The first-order rate constant for the activation process in the assay mixture is ATP-dependent at any level of inhibitory ADP. The data obtained strongly suggest that two ADP-specific inhibitory sites and one ATP-specific hydrolytic site are present in F₁-F₀ ATPase. Taking into account the (3α·3β)·γ·δ·ϵ structure of F₁, it is concluded that the synchronous discharge of ADP from two inhibitory sites during the activation occurs after ATP binds to the ATPase catalytic site.

【 授权许可】

Unknown